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| Status |
Public on Feb 28, 2022 |
| Title |
scRNAseq of co-culture of primary human oligodendrocytes and IL-23 polarized human CD4 T cells in direct contact (contact) versus separated by a porous membrane (insert, no contact) |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Multiple sclerosis (MS) is characterized by the loss of myelin and of myelin-producing oligodendrocytes (OLs) in the central nervous system (CNS). Pro-inflammatory CD4+ Th17 cells are considered pathogenic in MS and are harmful to OLs. We investigated the mechanisms driving human CD4+ T cell-mediated OL cell death. Using single cell RNA sequencing, we assessed the transcriptomic profile of primary human OLs and Th17 cells in direct contact or separated by an insert. We showed that upon close interaction, OLs upregulate the expression of mRNA coding for chemokines and antioxidant/anti-apoptotic molecules, while Th17 cells upregulate the expression of mRNA coding for chemokines and pro-inflammatory cytokines such as IL-17A, IFN-γ and granzyme B. We found that secretion of CCL3, CXCL10, IFN-γ, TNFα and granzyme B is induced upon direct contact in co-cultures of human Th17 cells with human OLs. In addition, we validated by flow cytometry and immunofluorescence that granzyme B levels are upregulated in Th17 compared to Th2 cells, and are even higher in Th17 cells upon direct contact with OLs or MO3.13 cells compared to Th17 cells separated from OLs by an insert.
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| Overall design |
Adult human oligodendrocytes in primary culture derived from the same brain sample (resection path of surgery for epilepsy) were exposed to IL-23-polarized CD4 T cells (Th17 cells) from the same donor at 1:1 ratio. They were either directly in contact (contact condition) or separated by a an insert with a porous membrane of 0.4 µm size (insert, no contact condition).
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| Contributor(s) |
Larochelle C, Stratton J, Antel J, Cui Q, Jamann H, Cui Q |
| Citation(s) |
35479072 |
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| Submission date |
Feb 17, 2022 |
| Last update date |
May 05, 2022 |
| Contact name |
Olivier Tastet |
| E-mail(s) |
o.tastet33@gmail.com
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| Organization name |
crCHUM
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| Street address |
900 Saint-Denis
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| City |
Montreal |
| ZIP/Postal code |
H2X0A9 |
| Country |
Canada |
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| Platforms (1) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
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| Samples (2) |
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| Relations |
| BioProject |
PRJNA808071 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE196953_RAW.tar |
28.9 Mb |
(http)(custom) |
TAR (of CSV) |
| GSE196953_cell_metadata.csv.gz |
199.4 Kb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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